Excess rNTPs added 10 15 Time, minutes Incorporation of radioactivity, cpm

Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
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A mixture of four a-[32P]–labeled ribonucleoside triphosphates was added to permeabilized bacterial cells undergoing DNA replication in the presence of an RNA polymerase inhibitor, and incorporation into high-molecular-weight material was followed over time, as shown in the accompanying graph. After 10 minutes of incubation, a 1000-fold excess of unlabeled ribonucleoside triphosphates was added, with the results shown in the graph.

(a) Why was the excess of unlabeled rNTPs added?
(b) How could you tell that radioactivity is being incorporated as ribonucleotides rather than as an alternative such as reduction to deoxyribonucleotides, followed by incorporation?
(c) What does this experiment tell you about the process of DNA replication?

Excess
rNTPs
added
10
15
Time, minutes
Incorporation of radioactivity, cpm
Transcribed Image Text:Excess rNTPs added 10 15 Time, minutes Incorporation of radioactivity, cpm
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